KR20180037930A - Composition for Treatment of Pancreatic CancerComprising Extract of Myristicae Semen - Google Patents

Abstract

The present invention relates to a new use of a myristicae semen extract. More specifically, the present invention relates to a composition for treatment containing a myristicae semen extract as an active ingredient which represents excellent prevention or treatment effects for a pancreatic cancer. The composition of the present invention contains the myristicae semen extract as the active ingredient, additionally includes a pharmaceutically acceptable carrier or a diluent.

Description

Technical Field [0001] The present invention relates to a composition for treating pancreatic cancer,

The present invention relates to a novel use of nutmeg extract. Specifically, the present invention relates to a therapeutic composition comprising, as an active ingredient, a nutmeg extract exhibiting excellent preventive or therapeutic efficacy against pancreatic cancer.

Among the major diseases of modern people, studies on the methods of treatment and diagnosis of cancer are progressing relatively actively, focusing on lung cancer, liver cancer, stomach cancer which have high incidence. However, studies on esophageal cancer, colorectal cancer, and pancreatic cancer which have low incidence are relatively low.

The pancreas produces insulin that helps convert glucose into energy in the body and enzymes that help digest food in the body. Pancreatic cancer is a malignant growth of the pancreas, mainly occurring in cells of the pancreatic duct. The disease is the ninth most common form of cancer, but it is the fourth and fifth most common cause of cancer deaths for men and women, respectively. Pancreatic cancer has a 5-year survival rate of less than 3%, and most are always fatal. Among pancreatic cancer, pancreatic ductal adenocarcinoma (pancreatic ductal adenocarcinoma) in the pancreatic duct accounts for more than 90%. Generally, pancreatic cancer refers to pancreatic ductal adenocarcinoma. Pancreatic duct adenocarcinoma is also called pancreatic adenocarcinoma, pancreatic adenocarcinoma or pancreatic ductal cancer.

 Like many other malignant diseases, pancreatic cancer arises from the accumulation of acquired mutations. Multiple genetic and epigenetic changes, including activation of protooncogenes, inactivation of tumor suppressor genes, and malformations of maintenance genes have been linked to the development, persistent growth, and metastasis of pancreatic cancer . Accumulated mutations in these genes are known to occur within a predictable period of time during the "PanINs" (Pancreatic Intraepithelial Neoplasia) stage (Hruban et al. (2000) Clin Cancer Res 6: 2969-2972; Kern et al. (2002) Cancer Biol Therapy 1: 607-613; Li et al. (2004) Lancet 363: 1049-1057). The mutation of K-ras occurs at about half of the PanIN-1. The PanIN-2 stage is indicated by an additional change and an increase in the K-ras mutation rate, and a plurality of p16 malformations, and p53 protein family expression, which may indicate the presence of p53 mutations, is occasionally seen in more advanced PanINs. Loss of tumor suppressor genes, TP53, DPC4 and BRCA2, appears to occur later in the development of pancreatic tumor formation, PanIN-3. Specifically, over 85% of pancreatic ductal adenocarcinomas have a K-ras gene point mutation activated in the development of pancreatic cancer (Li et al. (2004) Lancet 363: 1049-1057; Xiong Pharm 54: S69-77). The K-ras mutation induces cell proliferation and activates the constitutive elements of Ras-Raf-MEK-ERK, an intracellular signaling pathway that allows this gene to be transfected with cells that contain point mutations . The ras mutation is not associated with the tumor stage or prognosis and indicates that the K-ras oncogene may be involved in the onset of carcinogenesis but is not associated with the possibility or promotion of malignancy of human pancreatic cancer. One of the major downstream targets of the ras family is phosphoinositol 3 kinase (PI3K). Activation of PI3K is associated with pancreatic cancer resistance to apoptosis induced by chemotherapy or molecular drug targeting agents.

Pancreatic duct adenocarcinoma is one of the most deadly human malignant tumors in the United States, with more than 30,000 deaths annually. At the time of diagnosis, these cancers are found to be only able to be resected, with only 10% to 15% being due to the presence of locally advanced disease or distant metastasis. Recently, the most common treatment strategy for advanced pancreatic cancer is gemcitabine, an intravenous 2'-deoxycytidine neuroleoside analogue that can induce apoptosis of human pancreatic cancer cells and inhibit tumor growth and progression, Lt; / RTI > However, despite the best medical or surgical treatment, the outcome of patients with pancreatic ductal adenocarcinoma is dismal and, as a group, the median survival of patients with this disease is less than 21 months. Therefore, clear, new and effective treatment strategies are needed to combat this fatal disease. Thus, pancreatic cancer is a major health issue in developed countries and has a very poor prognosis (Faint et al. (2004) Datamonitor DMHC2045; Garcea et al. (2005) Pancreatology 5: 514-529; Kern et al. Cancer Biol Therapy 1: 607-613; Laheru and Jaffee (2005) Nature Rev Cancer 5: 59-467; Li et al. (2004) Lancet 363: 1049-1057). Despite excessive surgical and medical treatment, the average life expectancy is approximately 15-18 months for patients with local disease and 3-6 months for patients with metastatic disease. Nearly 100% of patients with pancreatic cancer experience metastasis and die due to their ability to weaken metabolism by their unrestrained growth, and the likelihood of survival for a total of five years without resection is only less than 5% Do. Also, there is a problem that early diagnosis is difficult due to initial symptoms that are not specific. Currently, early detection of pancreatic cancer is still under development and not commercially available, and conventional cancer therapy has little effect on prognosis or disease outcome. Poor prognosis of pancreatic cancer is due to late signs, aggressive local invasion, early metastasis and insufficient response to chemotherapy.

The most common symptoms of pancreatic cancer are jaundice, abdominal pain and weight loss, and are not really special with other adverse factors. Thus, in the early stages of tumor growth, diagnosing pancreatic cancer is usually difficult, and requires considerable suspicion and extensive diagnostic overcrowding, often involving exploratory surgery. Endoscopic ultrasonography and computed tomography (CT) are currently the best noninvasive methods available for the diagnosis of pancreatic cancer. However, definite detection of small tumors as well as differentiation of pancreatic cancer from lesion pancreatitis is difficult. Unfortunately, the majority of patients are now diagnosed at the terminal stage where the tumor has already invaded the surrounding organs and extended outside the caecum and / or at the extensively metastasized stage (Gold et al., Crit. Rev. Oncology / Hematology, 39: 147-54 (2001)). The disease is delayed detection is common, and 'early' pancreatic cancer diagnosis is rare in the clinical setting.

Current treatments that can be used for pancreatic cancer do not reach the healing or substantially improved survival time of the disease. Surgical resection is the only way to provide an opportunity for survival. However, due to tumor burden, only 10-25% of patients are targeted for 'curative resection'. In these patients who experience resection, the 5-year survival rate is still on the order of 10% on average.

On the other hand, Myristicae Semen ( Myristica fragrans ) is an evergreen tree belonging to the Myristicaceae, which is obtained by picking off the seeds, drying the seeds, then removing the black hard seeds, dipping them overnight in the lime juice, and drying them at room temperature. Malaysia, Indonesia and Sri Lanka are the main producing countries and they are usually collected from April to June or November to December, and have been used to treat blood circulation, respiratory disorders and skin diseases by facilitating blood circulation and lung function and sterilizing effect It is also used as a fragrance. Studies on nutmeg have been reported on cytotoxic effects, antibacterial and antioxidative and antimicrobial effects on cancer cell lines, hepatotoxic effects, and brain collins teratase inhibition associated with dementia. Toxicol, 31: 517-21, 1993), the chemical activity of the skin warts (In Vivo, 17: 541-4, 2003) (Cancer Lett., 56: 59-63, 1991) and anti-inflammatory activity (Jpn. J. Pharmacol., 49: 155-63, 1999). Also, it is known that the herb medicine has converging action, branch action, textural action and gas discharge action.

Nutmeg is a seed of Myristica fragrans HOUTT. Its fruit is 4 ~ 6 ㎝ in length. When mature, it looks like an apricot, with seeds in it, and a reddish yellow bark on its inside, which shows the seeds of the cracks like the ribs. The seeds are called the nutmeg, and the seeds are called the mace. Breeding is mostly seed and grows well in high temperature and high humidity. Nutmeg is dried and used as a directional tablet, tonic, etc. It is also known to have antibacterial activity. The essential components include essential oil of 2 to 9%, d-camphene and alpha-pinene, and the content of tetradecanoic acid in fat is 70 to 80 % And myristin, which is a poisonous substance, is contained. It warms the stomach and spleen, and has efficacy on the following (气), news (erosion), and kite (kosang) , Young Lim, p. 59-120, 1998).

However, there has not yet been disclosed or disclosed in the above-mentioned literature regarding pancreatic cancer-related diseases of nutmeg.

Accordingly, the inventors of the present invention have completed the present invention by confirming that the nutmeg extract can effectively kill pancreatic cancer cells while conducting the herbal medicine for nutmeg.

It is an object of the present invention to provide a composition for treating pancreatic cancer, which comprises a nutmeg extract as an active ingredient.

In order to achieve the above object, the present invention provides a composition for preventing and treating pancreatic cancer, which comprises an extract of Myristicae Semen ethanol as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for preventing and treating pancreatic cancer, which comprises an extract of Myristicae Semen ethanol as an active ingredient. The composition of the present invention comprises a nutmeg extract as an active ingredient and may further comprise a pharmaceutically acceptable carrier or diluent.

In the composition for preventing and treating pancreatic cancer according to the present invention, it is preferable that the ethanol extract is extracted at 50 ° C for 24 hours, and the ethanol extract is more preferably dried and concentrated under reduced pressure at 45 ° C, Most preferably 95%.

In addition, in the composition for preventing and treating pancreatic cancer of the present invention, the pancreatic cancer is preferably pancreatic ductal adenocarcinoma.

The nutmeg extract of the present invention can be applied to any part of the nutmeg, but is preferably extracted from the fruit. The extraction solution can be obtained by extraction with water or an organic solvent. Examples of the organic solvent include lower alcohol, acetone, chloroform, methylene chloride, ether, ethyl acetate, hexane and the like. As the lower alcohol, methanol, ethanol, propanol and butanol can be mentioned, and ethanol is most preferable.

Specifically, 1 to 5 times, preferably 3 times, 95% ethanol is added to the dried or powdered nutmeg and the mixture is heated at a temperature of 20 to 100 캜, preferably 40 to 60 캜 for 10 to 100 hours, 40 hours, more preferably 24 hours, and then filtered to produce an ethanol extract of nutmeg. Preferably, the extract is filtered, and the resulting filtrate is concentrated under reduced pressure. In the above extraction methods, the extraction process may be repeated twice or more as necessary, and the extract obtained after filtration may be lyophilized or dried under reduced pressure to a powder form.

The term "pharmaceutically acceptable carrier" is intended to encompass pharmaceutically acceptable substances, such as liquid or solid fillers, diluents, excipients or solvents, which serve to transport the active ingredient from one or more parts of the body to other organs or parts of the body , Composition or vehicle.

The composition for treating pancreatic cancer of the present invention may be prepared from a pharmaceutical agent by adding one or more pharmaceutically acceptable carriers together with the active ingredient. Such carriers include, but are not limited to, saline, buffered saline, water, glycerol, and ethanol, and any suitable formulation known in the art (Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA) .

The pharmaceutical composition of the present invention can be administered orally in clinical administration and can be used in the form of a general pharmaceutical preparation. In the case of pharmaceutical preparation, a filler, an extender, a binder, a wetting agent, a disintegrant, A surfactant, or the like. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. Common diluents such as water, In addition to liquids and paraffins, various excipients such as wetting agent sweetening agents, perfumes, preservatives and the like may be included.

In addition, the herbal medicine which may be added to the composition of the present invention may be any pharmaceutically acceptable herbal medicine, for example, Angelicae tenuissimae Radix, Gastrodiae Rhizoma, Bapleuri Radix, (Rhizoma), Angelicae gigantis Radix, Persicae Semen, Cinnamomi Ramulus, Rhei Rhizoma, Glycyrrhizae Radix, Cnidii Rhizoma, Aurantii nobilis Pericarpium, Alismatis Rhizoma, Coptidis Rhizoma, Scutellariae Radix, Hoelen, Paeoniae Radix, Atractylodis Rhizoma alba, Phellodendri Cortex, Gardeniae Fructus, Pinelliae Tuber, Uncaria Ramuluset, Uncus, Ponciri Fructus, Ginseng, Liriopis Tuber, Polygalae Radix, Acori graminei Rhizoma, Atractylodis Rhizoma alba, Chrysanthemi Flos, Ledebouriellae Radix, ), Ginger (Zingiberis Rhizoma crudus), ganoderma (Natrii sulfas), control yphi Fructus, Salviae Radix, Mautan Radicis Cortex, Rehmanniae Radix, Menthae Herba, Dioscoreae Rhizoma, Polyporus, Polygonimultiflori Radix, Allii tuberosi Semen, Cassiae Semen, Lycii Fructus, Araliae cordatae Radix, Eucommiae Cortex, Hedyotis Herba, Saururus Herba, Artemisiaecapillaris Herba, Anemarrhenae, Rhizoma, Rhizoma, Carthami Flos, Astragali Radix, Lycopodium, Ginkgonis Folium, Polygonati Rhizoma, Nelumbinis Semen, Fossilia ossis Mastodi, Lycii radicis Cortex ), Achyranthis Radix, Rehmanniae Radix preparata, Perillae Semen, Thujae Semen, Hordei Fructus germinatus, Cuscutae Semen, Morindae Radix, Pini koraiensis Radix) may be used alone or in combination.

The composition of the present invention can be administered in various forms of parenteral administration at the time of actual clinical administration. Solid formulations include tablets, pills, powders, granules, capsules and the like. Examples of the liquid formulations include suspensions, , Syrups, and the like. Various excipients such as wetting agent sweeteners, fragrances, preservatives, etc. may be included in addition to simple diluents commonly used in water, liquids and paraffins. Specifically, formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. Calcium or vitamin D ₃ may also be added to enhance the efficacy of the therapeutic agent. Such compositions may be presented in unit-dose (one-time) or multi-dose (several-dose) containers, such as sealed ampoules and vials, and may be presented in a sterile liquid carrier, Lt; RTI ID = 0.0 > freeze-drying < / RTI > Immediate injectable solutions and suspensions may be prepared from sterile powders, granules and tablets.

The formulations of the present invention may be applied differently depending on the age, sex, condition of the subject, the degree of absorption of the active ingredient in the body, the rate of inactivation and the rate of excretion, and the drugs used in combination. The present invention also includes formulations of dosage units. The formulations are presented in separate dosage forms, such as tablets, coated tablets, capsules, pills, suppositories, and ampoules, and the content of active compound in the drug is a fraction or multiple of the individual dosage. Dosage units may contain, for example, 1, 2, 3 or 4 times, or 1/2, 1/3 or 1/4 times the individual doses. The individual doses preferably contain amounts in which the active compound is administered in a single dose, which usually corresponds to the full, half, one-third or one-fourth of the daily dose.

As used herein, the term "extract " means an active ingredient isolated from a natural product. The extract can be obtained by an extraction process using water, an organic solvent, or a mixed solvent thereof, and includes an extract, a dry powder thereof, or all the forms formulated with it.

In a specific embodiment of the present invention, the ethanol extract of nutmeg killed 85.5% of Panc-1 pancreatic cancer cells at 100 μg / ml, and the IC ₅₀ (half maximal inhibitory concentration) was 94.1 μg / ml. The above results demonstrate that the nutmeg extract of the present invention has an excellent killing activity of Panc-1 pancreatic cancer cells and further has a pancreatic cancer treatment and prevention activity.

As used herein, the term "prevention" means any action that inhibits or delays the onset of pancreatic cancer by administration of the composition.

In the present invention, the term "treatment" means any action that improves or alters the symptoms of pancreatic cancer by administration of the composition.

In the present invention, the nutmeg extract may be extracted by using water, an organic solvent, or a mixed solvent thereof. Preferably an organic solvent, especially ethanol. The extracted liquid can be used directly or by concentrating and / or drying. In the case of extraction using an organic solvent, an organic solvent such as methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF) (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof. The active ingredient of the herbal medicine may be extracted at room temperature or warmed under the condition that the active ingredient is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and the degree of loss of the active ingredient of the medicament may differ. Therefore, an appropriate organic solvent should be selected and used. The extraction method is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction. Filtration is a process of removing suspended solid particles from an extract, and may be performed by filtering particles using a cotton, nylon, or the like, or by ultrafiltration, freezing filtration, centrifugation, and the like.

Concentration of the extract can be carried out by reduced-pressure concentration, reverse osmosis concentration, or the like. The post-concentration drying step includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, infrared drying and the like. In some cases, a step of pulverizing the final dried extract may be added.

In addition, the extract may be subjected to an additional fractionation process. Preferably, the extract is suspended in distilled water, and the non-polar solvent soluble layer is extracted and separated by using a nonpolar organic solvent such as hexane, ether, dichloromethane, chloroform, ethyl acetate, or a mixed solvent thereof , Which can be used by concentration and / or drying.

In the present invention, the term "pharmaceutically acceptable salt" means salts derived from pharmacologically or physiologically acceptable inorganic acids, organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene- Formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals, such as sodium, alkaline earth metals, such as magnesium, ammonium, and the like.

The pharmaceutical composition for the prevention and treatment of pancreatic cancer diseases according to the present invention comprises 0.1 to 50% by weight of the above extract or compound, based on the total weight of the composition. In addition, the composition does not increase the efficacy, but may contain additional ingredients that are commonly used in pharmaceutical compositions to improve odor, taste, visual appearance, and the like. In addition, the composition may further include inorganic and organic additives such as vitamins B1, B2, B6, C, E, niacin, carnitine, betaine, folic acid pantothenic acid, biotin, zinc, iron, calcium, chromium, magnesium, As shown in FIG. In addition, the composition may include a substance having therapeutic activity against pancreatic cancer alone or in a previously used form.

The term "patient" in the present invention refers to a disease caused by pancreatic cancer and its direct or indirect cause, and can be administered to humans, horses, sheep, pigs, goats, camels, Nutrition, and dogs. By administering the composition comprising the nutmeg extract of the present invention to a patient, the above-mentioned pancreatic cancer can be effectively prevented and treated. The composition of the present invention can be administered in parallel with the existing pancreatic cancer therapeutic agent.

The term "administering" in the present invention means introducing a predetermined substance into a patient in any appropriate manner, and the administration route of the composition of the present invention may be oral or parenteral ≪ / RTI > The composition may also be administered by any device capable of transferring the active agent to the target cell.

The composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the sex of the patient, age, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. May be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art. The method for administration of the composition comprising the extract or the compound prepared according to the preparation method of the present invention is preferably oral or intravenous. Generally, the effective dose is usually 1 to 10 times, 500 mg / kg is preferable, and in case of intravenous administration, 1 to 100 mg / kg is preferable, and it can be administered 2-3 times a day. The dosage level for a particular patient can be varied depending on the sex, age, health condition, diet, time of administration, method of administration, drug mix, patient condition, and severity of neurological disease.

The present invention also provides a cosmetic composition for the prevention of pancreatic cancer comprising, as an active ingredient, an extract of Myristicae Semen ethanol containing a cosmetically acceptable cosmetic auxiliary additive.

The cosmetic composition of the present invention preferably contains 0.01 to 10% by weight, more preferably 0.1 to 1% by weight, of the ethanol extract in the total weight. The cosmetic composition of the present invention is not particularly limited in its formulation and may have cosmetic formulations of softening agent, nutritional lotion, massage cream, nutritional cream, pack, gel or skin adhesive type, Ointments, gels, creams, patches or spray formulations.

In the present invention, when the nutmeg extract of the present invention was injected into Panc-1 pancreatic cancer cells, it was confirmed that Panc-1 cells died (see FIG. 1 ). Thus, it is newly found that the extract of nutmeg has an effect of treating pancreatic cancer.

As described above, the nutmeg extract of the present invention inhibits the growth of pancreatic cancer cells and induces apoptosis. Therefore, the composition for treating pancreatic cancer according to the present invention will be very effective for the treatment of pancreatic cancer patients.

FIG. 1 shows the results of Alamar Blue assay to examine the effect of nutmeg on the growth of pancreatic cancer cells in human pancreatic cancer cells, Panc-1 cells, wherein the X axis represents the concentration of the nutmeg extract and the Y axis represents the survival of human pancreatic cancer cells Survival rate.

Hereinafter, the present invention will be described in more detail based on the following examples. It should be noted, however, that the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The present invention is not limited to the following examples. Will be apparent to those skilled in the art to which the present invention pertains.

Example 1: Preparation of nutmeg extract

3 kg of nutmeg (Chinese origin) purchased from Seoul Medicinal Plant was dried and crushed for 5 days at the shade and room temperature. The ground nutmeg was immersed in 30 L of 95% ethanol and extracted at 50 DEG C for 24 hours. This was filtered through a filter paper, dried and concentrated under reduced pressure at 45 캜 to obtain 278 g of a total extract, which was stored at -20 캜.

<Example 2> Effect of nutmeg extract on growth of pancreatic cancer cells

To investigate the effect of the extract of the nutmeg extract of Example 1 on the growth of pancreatic cancer cells, Panc-1 cells, a human pancreatic cancer cell, were treated with ethanol extract of nutmeg for 48 hours and analyzed by Alamar Blue. The Alamar Blue assay is a modified form of MTT assay that measures the fluorescence intensity of a product after degradation of a compound that is degraded by a specific enzyme into a living cell to determine the relative number of living cells after treatment It is an experimental method. This will be described in more detail below.

<2-1> Preparation and treatment of human pancreatic cancer cell line

Panc-1 cells, the pancreatic cancer cell line used in the present invention, were purchased from Korean Cell Line Bank (KCLB) and used for the experiment. Specifically, Panc-1 pancreatic cancer cells were cultured in Dulbeco's Modified Eagle's (DMEM) medium supplemented with 10% fetal bovine serum (Welgene) and 25 mM HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid) Medium) medium.

<2-2> Inhibition of Cell Growth of Panc-1 Pancreatic Cancer Cells

The extract of the nutmeg extract of Example 1 was found to inhibit the growth of pancreatic cancer cells, Panc-1 cells. Specifically, 4.5 X 10 ³ Panc-1 pancreatic cancer cells were seeded on a 96-well plate and cultured for 24 hours. Then, the nutrient broth (DMC) dissolved in DMSO Ethanol extract was treated for 48 hours at a concentration of 0 to 100 μg / ml (specifically, 0, 3.125, 6.25, 12.5, 25, 50 and 100 μg / ml, respectively) (Table 1). After treating each concentration of the extract, 20 μl of Alamar Blue reagent was added to 0.2 ml of the cell culture solution filled in each well in a 96-well plate, and the plate was cultured in an incubator for 2 hours. The plate was gently shaken to react cells in each well, and the intensity of fluorescence was measured at 590 nm using a fluorescence photometer (Molecular Devices Corp.) while irradiating light at a wavelength of 544 nm. Survival rate is shown in Fig.

Nutmeg concentration Pancreatic cancer cell survival (average) Standard Deviation 0 [mu] g / ml 1,000 0.000 3.125 [mu] g / ml 1.118 0.034 6.25 [mu] g / ml 1.159 0.057 12.5 占 퐂 / ml 1.146 0.045 25 [mu] g / ml 1.014 0.044 50 [mu] g / ml 1.026 0.072 100 [mu] g / ml 0.145 0.003

As a result, as shown in Table 1 and FIG. 1, the growth of pancreatic cancer cells was decreased as the treatment concentration of nutmeg was higher, suggesting that the nutmeg had a pancreatic cancer therapeutic effect. That is, pancreatic cancer cells were killed by 85.5% at 100 μg / ml. In addition, IC ₅₀ (half maximal inhibitory concentration) was measured at 94.1 / / ml. Table 1 shows the relative number of cells of pancreatic cancer cells after 48 hours according to the concentration of each nutmeg treatment on the basis of the number of survival rate of pancreatic cancer cells in the control group not treated with nutmeg. Thus, the nutmeg extract of the present invention has excellent Panc-1 cell killing activity, further demonstrating that it has pancreatic cancer treatment and prevention activity.

<Example 3> Acute toxicity test using nutmeg extract

The nutmeg used in the present invention was widely used as a medicinal agent, so it was considered that there would be no problem in stability. However, toxicity test was conducted by oral administration and intraperitoneal administration to confirm this.

Acute toxicity tests were carried out using 6-week-old SPF SD rats. The nutmeg extract of Example 1 of the present invention was suspended in a 0.5% methylcellulose solution and then administered orally at a dose of 5 g / kg in each of two animals per group. After the administration of the test substance, the mortality, clinical symptoms, and weight changes of the animals were observed, and hematological tests and blood biochemical tests were carried out, and autopsy was performed to observe the abnormalities of the abdominal organs and thoracic organs.

As a result of the test, there were no clinically symptomatic or dead animals in all the animals to which the test substance was administered, and no toxic change was observed in weight change, blood test, blood biochemical test, and autopsy findings. These results nutmeg extract does not represent a change in toxicity both to 5 g / ㎏ in rats administered oral minimum lethal dose (LD ₅₀₎ was determined to be a safe substance less than 5 g / ㎏.

<Preparation Example 1> Preparation of therapeutic agent for pancreatic cancer containing nutmeg extract as an active ingredient

The inventors of the present invention confirmed that the nutmeg extract had excellent efficacy in treating pancreatic cancer, and thus, a therapeutic agent for pancreatic cancer containing the extract as an active ingredient was prepared as follows. The preparation examples of the following therapeutic agents can be applied not only to therapeutic agents but also to the production of cosmetic compositions.

&Lt; 1-1 > Soft gelatin capsules containing nutmeg extract (wt%)

Nutmeg extract 20%

Vitamin C 4.5%

Vitamin D ₃ 0.001%

Manganese sulfate 0.1%

Wax 10%

palm oil 25%

Safflower oil 30.399%

 &Lt; 1-2 > Preparation of an intravenous formulation containing nutmeg extract (% by weight)

Nutmeg extract 0.2%

Mannitol 0.3%

Saline solution 9.5%

&Lt; 1-3 > Tablets containing nutmeg extract (% by weight)

Nutmeg extract 35%

Vitamin C 10%

Vitamin D ₃ 0.001%

Manganese sulfate 0.1%

Crystalline cellulose 25.0%

Lactose 17.999%

Magnesium stearate 2%

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the invention, and are not intended to limit the scope of the invention. .

Claims (2)

A pharmaceutical composition for preventing or treating pancreatic cancer, comprising an organic solvent extract of nutmeg as an active ingredient. 2. The composition of claim 1, wherein the composition comprises a pharmaceutically acceptable carrier or diluent.

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